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Genetic mapping, cloning, and functional characterization of the BnaX.VTE4 gene encoding a γ-tocopherol methyltransferase from oilseed rape

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Abstract

Rapeseed (Brassica napus L.) is one of the major oilseed crops and an important source for tocopherols known as vitamin E in human nutrition. Increasing the tocopherol content and altering the tocopherol composition is a major goal in rapeseed breeding. The genes encoding enzymes from the tocopherol pathway have been cloned from model species. However, only scant data about tocopherol genes from crop species have been available. We have cloned four sequences of a gene family from B. napus with homology to the Arabidopsis thaliana VTE4 gene. The sequences were amplified by PCR with primers derived from the A. thaliana gene. BAC-clones were isolated to analyze the genomic structure of the BnaX.VTE4-loci. In contrast to the A. thaliana gene all B. napus sequences have two additional introns. For functional analysis, the BnaA.VTE4.a1 sequence was transformed into A. thaliana. Seeds from transgenic offspring showed a 50-fold increase of the α-tocopherol fraction which is in accordance with the predicted function of the gene. A marker assay was established and the BnaA.VTE4.a1 sequence was mapped to the end of chromosome A02 of the Tapidor × Ningyou7 genetic map, where also two QTL for α-tocopherol content had been mapped. Thus, the BnaA.VTE4.a1 gene is a promising candidate for these QTL and can be used for marker assisted selection for α-tocopherol content in rapeseed.

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Acknowledgments

We wish to thank Martina Bach and Jens Hermann for their excellent technical assistance. This work was funded by the Stiftung Schleswig-Holsteinische Landschaft and the German Research Foundation (DFG) as part of the Research Training Group GRK820.

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Authors and Affiliations

  1. Plant Breeding Institute, Christian Albrechts University of Kiel, Olshausenstr. 40, 24098, Kiel, Germany

    Jessica Endrigkeit & Christian Jung

  2. National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, 430070, Wuhan, People’s Republic of China

    Xingxing Wang, Chunyu Zhang, Yan Long & Jinling Meng

  3. Molecular Phytopathology, Christian Albrechts University of Kiel, Hermann-Rodewald-Str. 9, 24118, Kiel, Germany

    Daguang Cai

Authors
  1. Jessica Endrigkeit
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  2. Xingxing Wang
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  3. Daguang Cai
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  4. Chunyu Zhang
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  5. Yan Long
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  6. Jinling Meng
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  7. Christian Jung
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Corresponding author

Correspondence to Christian Jung.

Additional information

Communicated by H. Becker.

The nucleotide sequences data reported in this paper have been submitted to GenBank nucleotide sequence database with the accession numbers EU637012 to EU637015 and FJ435091 to FJ435093.

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122_2009_1066_MOESM1_ESM.ppt

S.1: Sequence alignment of the amplified regions using the primer combination vte4-4fw/vte4-4rv from three different BAC-derived BnaX.VTE4 sequences. Primer orientation is indicated by blue arrows. In red: deletion used to distinguish the BnaA.VTE4.a2 sequence from Tapidor from the other sequence variants by length polymorphism. Supplementary material 1 (PPT 105 kb)

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Endrigkeit, J., Wang, X., Cai, D. et al. Genetic mapping, cloning, and functional characterization of the BnaX.VTE4 gene encoding a γ-tocopherol methyltransferase from oilseed rape. Theor Appl Genet 119, 567–575 (2009). https://doi.org/10.1007/s00122-009-1066-6

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  • DOI: https://doi.org/10.1007/s00122-009-1066-6

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